DAG: Live Cell Diacylglycerol Assays
DAG Assay Performance
Off the shelf kits include the DAG sensor in BacMam, a BSL-1 viral vector for efficient delivery to most cell types. Purified BacMam, AAV, Lenti vectors by request.
- Easily detectable on plate readers or imaging systems
- Consistent well-to-well expression
- High signal-to-noise ratio
In HEK 293 cells over-expressing the P2Y11 receptor, ATP produces changes in both DAG and Ca2+, consistent with activation of Gq-coupled GPCR signaling. UTP produces no DAG response, indicating that the Ca2+ change is independent of Gq.
- Combine red and green sensors to measure multiple GPCR pathways simultaneously
- Red or green sensor version
- Single channel, non-FRET signal
- Minimal liquid handling
- No cell lysis
- 96 or 384 well compatible
- Herranz, Gonzalo et al. Protein Kinase C δ Regulates the Depletion of Actin at the Immunological Synapse Required for Polarized Exosome Secretion by T Cells. Frontiers in Immunology Apr. 2019
- Ohno, et al.Dynamics of Presynaptic Diacylglycerol in a Sensory Neuron Encode Differences between Past and Current Stimulus Intensity. Cell Reports, Vol. 20, Issue 10. Sept. 2017
- Shigeto, et al. GLP-1 stimulates insulin secretion by PKC-dependent TRPM4 and TRPM5 activation.J Clinical Invest. 2015. doi:10.1172/JCI81975.
- P. Rorsman, et al. GLP-1 stimulates insulin secretion by PKC-dependent TRPM4 and TRPM5 activation J Clinical Investigation Nov. 2015
- Arttamangkul, et al. Does PKC activation increase the homologous desensitization of μ opioid receptor? British Journal of Pharmacology.Jan. 2015.
- Xingjuan, et al. PKC-dependent Phosphorylation of the H1 Histamine Receptor Modulates TRPC6 Activity.Cells. 2014.
- P. Tewson, et al. New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories. J Biomol Screen Dec. 2015. Request Full Text PDF
- J. Williams, et al. Does PKC activation increase the homologous desensitization of μ opioid receptors? British Journal of Pharmacology April 2014
- Tewson, et al. Simultaneous Detection of Ca2+and Diacylglycerol Signaling in Living Cells.PLoS One. 2012.
- C. Peters, et al. Simultaneous detection of GPCR second messengers in living cells. BMG LabTech Dec. 2016.
“Assay is working beautifully with cardiomyocytes, showing a strong and immediate response.”
Expression in human islets is pretty superb, after 18 hours of incubation. An excellent probe!